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      BV605 Mouse Anti-Ki-67
      BV605 Mouse Anti-Ki-67
      Two-color flow cytometric analysis of Ki-67 expression by noncycling human peripheral blood mononuclear cells or proliferating MOLT-4 cells. Noncycling human peripheral blood mononuclear cells (PBMC; Top Plots) or proliferating cells from the human MOLT-4 (ATCC CRL-1582) cell line (Bottom Plots) were permeabilized and fixed with 70% ice-cold ethanol. The cells were washed twice with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), stained with either BD Horizon™ BV605 Mouse IgG1 Isotype Control (Cat. No. 562652; Left Plots) or BD Horizon™ BV605 Mouse Anti-Ki-67 antibody (Cat. No. 567122; Right Plots) and counterstained with BD Pharmingen™ DAPI Solution (Cat. No. 564907) to stain DNA. Bivariate pseudocolor density plots showing the correlated expression of DAPI staining versus Ki-67 (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact PBMC or MOLT-4 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
      BV605 Mouse Anti-Ki-67
      Two-color flow cytometric analysis of Ki-67 expression by noncycling Human peripheral blood mononuclear cells or proliferating MOLT-4 cells. Noncycling Human peripheral blood mononuclear cells (PBMC; Top Plots) or proliferating cells from the Human MOLT-4 (ATCC® CRL-1582™) cell line (Bottom Plots) were permeabilized and fixed with 70% ice-cold ethanol. The cells were washed twice with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), stained with either BD Horizon™ BV605 Mouse IgG1, κ Isotype Control (Cat. No. 562652; Left Plots) or BD Horizon™ BV605 Mouse Anti-Ki-67 antibody (Cat. No. 567122; Right Plots) and counterstained with BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) to stain DNA. Bivariate pseudocolor density plots showing the correlated expression of 7-AAD staining versus Ki-67 (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact PBMC or MOLT-4 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.
      BV605 Mouse Anti-Ki-67 BV605 Mouse Anti-Ki-67 BV605 Mouse Anti-Ki-67 BV605 Mouse Anti-Ki-67 BV605 Mouse Anti-Ki-67
      Two-color flow cytometric analysis of Ki-67 expression by noncycling human peripheral blood mononuclear cells or proliferating MOLT-4 cells. Noncycling human peripheral blood mononuclear cells (PBMC; Top Plots) or proliferating cells from the human MOLT-4 (ATCC CRL-1582) cell line (Bottom Plots) were permeabilized and fixed with 70% ice-cold ethanol. The cells were washed twice with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), stained with either BD Horizon™ BV605 Mouse IgG1 Isotype Control (Cat. No. 562652; Left Plots) or BD Horizon™ BV605 Mouse Anti-Ki-67 antibody (Cat. No. 567122; Right Plots) and counterstained with BD Pharmingen™ DAPI Solution (Cat. No. 564907) to stain DNA. Bivariate pseudocolor density plots showing the correlated expression of DAPI staining versus Ki-67 (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact PBMC or MOLT-4 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
      Two-color flow cytometric analysis of Ki-67 expression by noncycling Human peripheral blood mononuclear cells or proliferating MOLT-4 cells. Noncycling Human peripheral blood mononuclear cells (PBMC; Top Plots) or proliferating cells from the Human MOLT-4 (ATCC® CRL-1582™) cell line (Bottom Plots) were permeabilized and fixed with 70% ice-cold ethanol. The cells were washed twice with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), stained with either BD Horizon™ BV605 Mouse IgG1, κ Isotype Control (Cat. No. 562652; Left Plots) or BD Horizon™ BV605 Mouse Anti-Ki-67 antibody (Cat. No. 567122; Right Plots) and counterstained with BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) to stain DNA. Bivariate pseudocolor density plots showing the correlated expression of 7-AAD staining versus Ki-67 (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact PBMC or MOLT-4 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.
      Product Details
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      BD Horizon™
      MKI67; Antigen identified by monoclonal antibody Ki-67; KIA
      Human (QC Testing), Mouse (Tested in Development), Rat,Rhesus (Reported)
      Mouse IgG1, κ
      Human Ki-67
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      4288
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      Recommended Assay Procedures

         BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

         For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
      3. An isotype control should be used at the same concentration as the antibody of interest.
      4. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
      5. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      7. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
      8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      9. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
      10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      11. For U.S. patents that may apply, see bd.com/patents.
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      567122 Rev. 4
      Antibody Details
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      B56

      The B56 monoclonal antibody specifically binds to the Ki-67 antigen that is expressed in the nucleus of cycling cells (G1, S, G2, M cell cycle phases). During the G0 phase, the antigen cannot be detected. During interphase of the cell cycle, it is associated with nucleolar components, and it is on the surface of the chromosomes during M phase. Ki-67 is a large protein having 2 alternatively spliced isoforms, an N-terminal forkhead-associated domain, a C-terminal domain that binds to heterochromatin proteins, and multiple phosphorylation sites, the functions of which are still unclear. Because of the strict association of Ki-67 expression with cell proliferation, anti-Ki-67 antibodies are useful for the identification, quantification, and monitoring of growing cell populations.

      567122 Rev. 4
      Format Details
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      BV605
      The BD Horizon Brilliant Violet™ 605 (BV605) dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 407-nm and an acceptor dye with an emission maximum (Em Max) at 605-nm. BV605, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 610-nm (e.g., a 610/20-nm bandpass filter). The acceptor dye can be excited by the yellow-green (561-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BV605
      Violet 405 nm
      407 nm
      605 nm
      567122 Rev.4
      Citations & References
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      Development References (11)

      1. Benson MJ, Elgueta R, Schpero W, et al. Distinction of the memory B cell response to cognate antigen versus bystander inflammatory signals. J Exp Med. 2009; 206(9):2013-2025. (Clone-specific: Flow cytometry). View Reference
      2. Bigley V, Haniffa M, Doulatov S, et al. The human syndrome of dendritic cell, monocyte, B and NK lymphoid deficiency. J Exp Med. 2011; 208(2):227-234. (Clone-specific: Flow cytometry). View Reference
      3. Bruno S, Darzynkiewicz Z. Cell cycle dependent expression and stability of the nuclear protein detected by Ki-67 antibody in HL-60 cells. Cell Prolif. 1992; 25(1):31-40. (Biology: Flow cytometry). View Reference
      4. Kill IR. Localisation of the Ki-67 antigen within the nucleolus: evidence for a fibrillarin-deficient region of the dense fibrillar component. J Cell Sci. 1996; 109(6):1253-1263. (Biology). View Reference
      5. Kouro T, Medina KL, Oritani K, Kincade PW. Characteristics of early murine B-lymphocyte precursors and their direct sensitivity to negative regulators. Blood. 2001; 97(9):2708-2715. (Clone-specific: Flow cytometry). View Reference
      6. Kubbutat MH, Key G, Duchrow M, Schluter C, Flad HD, Gerdes J. Epitope analysis of antibodies recognising the cell proliferation associated nuclear antigen previously defined by the antibody Ki-67 (Ki-67 protein). J Clin Pathol. 1994; 47(6):524-528. (Biology). View Reference
      7. Picker LJ, Hagen SI, Lum R, et al. Insufficient production and tissue delivery of CD4+ memory T cells in rapidly progressive simian immunodeficiency virus infection. J Exp Med. 2004; 200(10):1299-1314. (Clone-specific: Flow cytometry). View Reference
      8. Pitcher CJ, Hagen SI, Walker JM, et al. Development and homeostasis of T cell memory in rhesus macaque. J Immunol. 2002; 168(1):29-43. (Clone-specific: Flow cytometry). View Reference
      9. Scholzen T, Gerdes J. The Ki-67 protein: from the known and the unknown.. J Cell Physiol. 2000; 182(3):311-22. (Biology). View Reference
      10. Spargo LDJ, Cleland LG, Cockshell MP, Mayrhofer Graham. Recruitment and proliferation of CD4+ T cells in synovium following adoptive transfer of adjuvant-induced arthritis. Int Immunol. 2006; 18(6):897-910. (Clone-specific: Flow cytometry, Immunofluorescence).
      11. Valenti LM, Mathieu J, Chancerelle Y, et al. High levels of endogenous nitric oxide produced after burn injury in rats arrest activated T lymphocytes in the first G1 phase of the cell cycle and then induce their apoptosis. Exp Cell Res. 2005; 306(1):150-167. (Clone-specific: Flow cytometry). View Reference
      View All (11) View Less
      567122 Rev. 4

      Please refer to Support Documents for Quality Certificates


      Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


      Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

      For Research Use Only. Not for use in diagnostic or therapeutic procedures.

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